T. G. Ohm

Close-meshed prevalence rates of different stages as a tool to uncover the rate of Alzheimer's disease-related neurofibrillary changes

The speed of progression of Alzheimers disease-related neurofibrillary changes is unknown. One reason for this is the impossibility to histopathologically follow-up one and the same individual over decades of their life. The present approach takes advantage of a recently introduced classification system which allows for a ranking of Alzheimers disease-related neurofibrillary changes into six stages [Braak and Braak Acta Neuropath (1991) 82, 239-259] and analyses a staged sample of 887 brains obtained from routine autopsy. It sets out to interpret these cross-sectional data in dynamic longitudinal terms, in order to estimate the rate of passing through the various stages. The time needed to attain respective stages of pathology for 5% of a given cumulative sample is determined. The resulting fifth centiles are a measure of the average rate by which the disease-related changes progress assuming that the underlying stages represent a sequence of events and do not independently emerge. Advancing age and the prevalence of Alzheimers disease-related changes of a given stage show a nonlinear positive correlation with only slight acceleration above the age of 65 years. Statistically, it takes at least 16 years from stage I to stage II, about 14 years pass by from stage II to III, 13 years from stage III to IV and five years from stage IV to V (= Alzheimers disease) for 5% of a given cumulative sample. Thus, the deep roots of Alzheimers disease-related neurofibrillary changes can be traced about 50 years back and may even extend into adolescence.(ABSTRACT TRUNCATED AT 250 WORDS)

Apolipoprotein E polymorphism influences not only cerebral senile plaque load but also Alzheimer-type neurofibrillary tangle formation

Only recently, evidence was provided that apolipoprotein E allele epsilon 4 located on Chromosome 19 is associated with late onset (i.e. senile) sporadic Alzheimers disease. Histologically, Alzheimers disease is associated with intraneuronal neurofibrillary changes and extraneuronal A4/beta-amyloid deposition. We set out with a histological staging system which considers the gradual development of Alzheimers disease-related histological changes over time and correlates highly with the cognitive decline ante mortem. Our analysis revealed that both the mean stage for A4/beta-amyloid deposits and the mean stage for neurofibrillary tangles get significantly shifted upwards in epsilon 4-carriers. This represents an earlier onset of the histopathological process of about one decade. The fact that both types of Alzheimers disease-related changes correlate positively with the prevalence of the epsilon 4-allele suggests for a causal relationship between the apolipoprotein E polymorphism and the development of Alzheimers disease.

Silver Impregnation of Alzheimer's Neurofibrillary Changes Counterstained for Basophilic Material and Lipofuscin Pigment

A method is described in which selective silver staining of Alzheimers neurofibrillary changes is combined with staining of cell nuclei, Nissl material, and lipofuscin granules. Formalin fixed, paraffin embedded sections of human autopsy tissue are silver stained according to a method proposed by Gallyas. Lipofuscin is stained by crotonaldehyde fuchsin following performic acid oxidation. Nissl substance is visualized by either Darrow red or gallocyanin-chrome alum staining. Architectonic units showing the specific pathology and the neuronal types prone to develop the neurofibrillary changes can be recognized using this technique.

Reduced basal and stimulated (isoprenaline, Gpp(NH)p, forskolin) adenylate cyclase activity in Alzheimer's disease correlated with histopathological changes

Cyclic adenosine monophosphate (cAMP) is an adenylate cyclase borne second messenger involved in basic metabolic events. The beta-adrenoceptor sensitive adenylate cyclase was studied in post-mortem hippocampi of controls and Alzheimer patients. Virtually identical subsets of each hippocampus homogenate were stimulated by 100 mumol isoprenaline, Gpp(NH)p and forskolin, respectively, in presence of an ATP-regenerating system. The determination of cAMP formed was carried out by means of a radioassay. The observed significant 50% reduction in basal as well as in stimulated adenylate cyclase activity in Alzheimers disease is negatively correlated with semiquantitative evaluations of amyloid plaques (P less than 0.05) but not with neuritic plaques, neurofibrillary tangles or neuropil threads. This reduction in enzyme activity is obviously not due to simple cell loss alone. It is likely that the crucial point of the observed functional disturbance is at the level of the catalytic unit of the adenylate cyclase, since the same degree of reduction is maintained at all steps of the signal cascade.

Chronopharmacokinetics of beta-receptor blocking drugs of different lipophilicity (Propranolol, metoprolol, sotalol, atenolol) in plasma and tissues after single and multiple dosing in the rat

SummaryComparative pharmacokinetic studies with the β-receptor blocking drugs propranolol, metoprolol, sotalol and atenolol, differing greatly in lipophilicity, and their main route of elimination were performed in light-dark-synchronized rats after equimolar single (6 μmoles/kg) or multiple (6x6 μmoles/kg) drug application. Drug concentrations were determined in plasma and various target organs of the drugs, e.g. heart, muscle, lung and brain, after drug application in the light period (L) and dark period (D), respectively. After single drug administration pharmacokinetic parameters of all drugs depended on the L and D conditions. Elimination half-lives in plasma and organs were shorter during D than during L. No L-D-differences were found in initial drug concentrations of the hydrophilic drugs sotalol and atenolol. In contrast, C0-values of the lipophilic propranolol in highly perfused organs (muscle, lung, brain) and of metoprolol in muscle tissue were significantly higher in D than in L. No obvious temporal dependency was found in other pharmacokinetic parameters (AUC, plasma clearance,Vdβ) with the exception inVdβ of propranolol. Due to the different physico-chemical properties of the compounds inter-drug-differences in pharmacokinetic parameters including drug accumulation into lung and brain tissue were observed. Multiple drug dosing abolished the circadian-phase-dependency in the elimination half-lives of the drugs due to an increase in D. Only for the highly lipophilic propranolol half-lives in highly perfused organs were still shorter in D than in L. It is concluded that L-D-differences in drug half-lives after single dose application are mainly due to circadian variations in drug elimination with a higher hepatic (propranolol, metoprolol) or renal (sotalol, atenolol) elimination in the activity period of rats during D. Additional studies with propranolol on heart rate of conscious rats revealed that a maximum in β-receptor blockade was achieved at 10 μmoles/kg in L but not in D. Thus, it is assumed that abolition of circadian-phase-dependency in half-lives after 6x6 μmoles/kg of the drugs may be due to the longer lasting and more pronounced β-receptor blockade after multiple drug dosing over a period of several hours in D. Thereby, liver-flow-dependent elimination of propranolol and metoprolol and renal elimination of sotalol and atenolol is reduced to base-line levels found in L.

Adenylyl cyclase activity in Alzheimer's disease brain : stimulatory and inhibitory signal transduction pathways are differently affected

Adenylyl cyclase (AC) activity was studied in post mortem hippocampus and cerebellum from eight patients with Alzheimers disease/senile dementia of the Alzheimer type (AD/SDAT) and seven non-demented control patients. AC was stimulated via stimulatory guanine nucleotide binding proteins (Gs) using guanosine triphosphate (GTP) and GppNHp (both 10(-4) M) or directly with either forskolin (10(-4) M) or Mn2+ (10(-2) M). Inhibition of AC via A1-receptors was performed with N6-cyclohexyladenosine (CHA) under basal conditions and in the presence of forskolin (10(-5) M). In both brain regions AC activity was significantly reduced in AD/SDAT when compared to controls. Under basal conditions and after stimulation via Gs mean reduction in hippocampus and cerebellum was 47.7% and 58.2%, respectively. The reduction was less pronounced after direct activation of the AC, amounting to 21.8% in hippocampus and 28.1% in cerebellum. CHA inhibited basal and forskolin-stimulated AC concentration-dependently by about 20% (basal) and 30% (forskolin). Inhibition by CHA was similar in hippocampus and cerebellum and tended to be more pronounced in AD/SDAT than in controls. Since the reduction of AC activity in AD/SDAT is greater after stimulation via Gs than after direct activation of the catalytic subunit, we suggest that both Gs and the catalytic subunit seem to be impaired. The fact that CHA-mediated inhibition of AC is not significantly different in AD/SDAT and controls, indicates that in contrast to Gs-, inhibitory G-proteins (Gi) coupling to AC remains intact in Alzheimers disease.

Neuropeptide Y-like immunoreactive neurons in the human olfactory bulb

Neuropeptide Y-like (NPY) immunoreactivity was localized in the adult human olfactory bulb by the unlabeled antibody enzyme (peroxidase anti-peroxidase; PAP) technique in vibratome sections. The majority of NPY-immunoreactive somata was localized in the white matter surrounding the anterior olfactory nucleus. Immunoreactive neurons were less numerous within the anterior olfactory nucleus and within the olfactory bulb layers. NPY-immunoreactive fibres were present in the white matter, the anterior olfactory nucleus, and in the olfactory bulb layers. Fibres within the white matter were generally aligned in a straight path parallel to the long axis of the olfactory bulb and tract. Fibres within the anterior olfactory nucleus showed no clear orientation and displayed numerous branching points. Coiled plexus of NPY-immunoreactive fibres were present in the glomerular layer of the olfactory bulb. Additional characteristics of the NPY-immunoreactive neurons were studied after decolouring the chromogen and restaining the sections with aldehydefuchsin to demonstrate the presence of lipofuscin granules and also with gallocyanin chrome alum to stain the Nissl substance. This analysis showed that the neurons belong to the class of non-pigmented nerve cells.

The organotypic entorhinal-hippocampal complex slice culture of adolescent rats. A model to study transcellular changes in a circuit particularly vulnerable in neurodegenerative disorders

The entorhinal-hippocampal system is severely altered in many neurodegenerative disorders with mnemonic malfunction, e.g. Alzheimers, Parkinsons and Huntingtons disease. The present approach characterizes an organotypic complex slice culture comprising both the entorhinal cortex and the hippocampal formation in order to establish a tool for experimental studies of the entorhinal-hippocampal interaction and its presumed neurodegenerative alterations in vitro. Slices were obtained from rats at about postnatal day 15 and maintained in culture using the interface technique. Thus, also structures known to be developed gradually during the first weeks postnatally are in accord to structures seen in adult rats. After two-three weeks in vitro, slices in the culture dish still revealed the typical morphological features of the entorhinal-hippocampal formation as visible with the dissecting microscope. Biocytin, which is taken up by and transported within living cells, labeled typical cell bodies, dendrites and axons of stellate neurons in layer II and pyramidal cells in layer III when applied to the outer layers of the entorhinal cortex. Small injections of biocytin within the dentate gyrus displayed living granule cells and the maintenance of their projection to the pyramidal cells in CA3, i.e., a typical suprapyramidal plexus of mossy fibers. The presence of axons of entorhinal neurons traveling towards the hippocampus and growth cones traversing the deep layers of the entorhinal cortex indicate that both brain regions are still interacting. Immunocytochemistry for calbindin D-28K revealed labeled neurons in layer II of the entorhinal cortex and dentate granule cells which are known to contain this calcium-binding protein.

Auditory brainstem nuclei in Alzheimer's disease

Serial sections of 7 Alzheimer brainstems were examined. The histopathological hallmarks were demonstrated by means of Kongo red- and silver-stained preparations. A subsequent counterstaining with a Nissl dye, allowing the cytoarchitectonical interpretation, revealed a considerable plaque formation in the central nucleus and the dorsomedial nucleus of the inferior colliculus. To a lesser degree plaques were also present in the deep layers of the dorsal cortex of the inferior colliculus. All other auditory brainstem nuclei were devoid of neuritic plaques. Neurofibrillary tangles were rarely seen. They occur in the dorsal cochlear nucleus, the periolivary region, the ventral nucleus of the lateral lemniscus, and in the central nucleus of the inferior colliculus.

Glutamine synthetase immunoreactivity in the human hippocampus is lamina-specific

Vibratome sections of immersion-fixed (4% paraformaldehyde) hippocampi from four autoptic cases without neuropathological findings were immunostained with a polyclonal anti-glutamine synthetase antibody. Immunoreactivity is restricted to astrocytes and has a layer-specific distribution: The highest levels are observed in the outer molecular layer of the fascia dentata, in the pyramidal cell layer and in the upper stratum oriens. The stratum radiatum displays particularly low immunoreaction. A conspicuous drop in immunoreactivity from the heavily stained outer molecular layer to its inner portion is marked by a clear-cut dividing line. The layer-specific staining pattern found corresponds to the laminated distribution of glutamate binding sites described by others.